Trial manufacture of an ultra-high sensitive video-microscope system with and image-processor and its applications to cell biology

带图像处理器的超高灵敏度视频显微镜系统的试制及其在细胞生物学中的应用

• 批准号: 59840020
• 负责人: HIRAMOTO Yukio
• 金额: $ 20.42万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1984
• 资助国家: 日本
• 起止时间: 1984 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-59840020/
• 关键词: Intracellular <Ca^(2+)>; aequorin; high-sensitive video-microscopy; fertilization; cleavage; amoeba; シラタマモ; 分裂周期; 顕微鏡; 高感度テレビ; 細胞内カルシウム

A video-microscope system was developed, in order to visualize microscopic objects under an extremely-low intensity light. Using this system, intracellular release of calcium ions at fertilization and artifical activation was recorded in eggs of sea urchin, starfish, medaka, frog and golden hamster, which had been injected with aequorin, a photoprotein sensitive to free calcium ions. In these eggs, the increase in intracellular calcium ion concentration was started at the site of sperm entry or the site of artificial activation and it propagated over the cell cortex toward the antipod. This calcium wave preceded the wave of the breakdown of cortical alveoli or cortical granules, suggesting that the increase in calcium ions induces the cortical change.Periodic changes in intracellular calcium ions accompanying cell cycle were recorded in fertilized medaka and sea urchin eggs injected with aequorin. In both eggs, intracellular calcium ion concentration attained a minimum during cleavage.Transient and localized increases in intracellular calcium ions accoumpanying amoevoid movement and protoplasmic streaming in slime mold were recorded using aequorin as a probe.Changes in intracellular calcium concentration were also recorded using fura 2, a fluorescent probe for calcium ions.

开发了一种视频显微镜系统，以便在极低强度的光下观察微观物体。使用该系统，在注射了水母发光蛋白（一种对游离钙离子敏感的发光蛋白）的海胆、海星、青鳉、青蛙和金仓鼠的卵中记录了受精和人工激活时细胞内钙离子的释放。在这些卵子中，细胞内钙离子浓度的增加开始于精子进入的部位或人工激活的部位，并在细胞皮层上向反足传播。这种钙波先于皮层肺泡或皮层颗粒的破裂波，表明钙离子的增加引起了皮层的变化。在注射水母发光蛋白的青鳉和海胆受精卵中记录到细胞内钙离子伴随细胞周期的周期性变化。在两种卵中，细胞内钙离子浓度在卵裂过程中均达到最低值，用水母发光蛋白（aequorin）作为探针，记录到细胞内钙离子浓度随黏菌的变形虫运动和原生质流而短暂和局部增加，用钙离子荧光探针fura 2也记录到细胞内钙离子浓度的变化。

项目成果

Yukio Hiramoto: Academic Press. Video-intensified microscopy as a technique for recording <Ca^(2+)> release in single cells. In "International Review of Cytology", (1987)

平本幸雄：学术出版社。

Y.Yoshimoto;T.Iwamatsu;K.Hirano;Y.Hiramoto: Development,Growth and Differentiation. 28. 583-596 (1986)

Y.Yoshimoto；T.Iwamatsu；K.Hirano；Y.Hiramoto：发展、成长和差异化。

S.Miyazaki;N.Hashimoto;Y.Yoshimoto;K.Kishimoto;Y.Igusa;Y.Hiramoto: Developmental Biology. 118. 259-267 (1986)

S.Miyazaki；N.Hashimoto；Y.Yoshimoto；K.Kishimoto；Y.Igusa；Y.Hiramoto：发育生物学。

Yasuaki Yoshimoto, Takasi Iwamatsu.and Yukio Hiramoto: "Cyclic changes in intracellular calcium levels associated with cleavage cycles in echinoderm and medaka eggs." Biomedical Research. 6. 387-394 (1985)

Yasuaki Yoshimoto、Takasi Iwamatsu 和 Yukio Hiramoto：“细胞内钙水平的周期性变化与棘皮动物和青鳉卵的卵裂周期相关。”

H.Y.Kubota,Y.Yoshimoto,M.Yoneda,Y.Hiramoto: Developmental Biology. 119. (1987)

H.Y.久保田，Y.吉本，M.米田，Y.平本：发育生物学。